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  • 1
    ISSN: 1432-1424
    Keywords: Na+, K+-pump ; Na+, K+-ATPase ; K+ content ; Epithelial cells ; Proliferation ; Attachment ; Ouabain ; Chloroquine ; Strophanthidin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Na+, K+-pumps of most eukaryotic animal cells bind ouabain with high affinity, stop pumping, and consequently loose K+, detach from each other and from the substrate, and die. Lack of affinity for the drug results in ouabain resistance. In this work, we report that Ma104 cells (epithelial from Rhesus monkey kidney) have a novel form of ouabain-resistance: they bind the drug with high affinity (Km about 4×10−8 m), they loose their K+ and stop proliferating but, in spite of these, up to 100% of the cells remain attached in 1.0 μm ouabain, and 53% in 1.0 mm. When 4 days later ouabain is removed from the culture medium, cells regain K+ and resume proliferation. Strophanthidin, a drug that attaches less firmly than ouabain, produces a similar phenomenon, but allows a considerably faster recovery. This reversal may be associated to the fact that, while in ouabain-sensitive MDCK cells Na+, K+-ATPases blocked by the drug are retrieved from the plasma membrane, those in Ma104 cells remain at the cell-cell border, as if they were cell-cell attaching molecules. Cycloheximide (10 μg/ml) and chloroquine (10 μm) impair this recovery, suggesting that it also depends on the synthesis and insertion of a crucial protein component, that may be different from the pump itself. Therefore ouabain resistance of Ma104 cells is not due to a lack of affinity for the drug, but to a failure of its Na+, K+-ATPases to detach from the plasma membrane in spite of being blocked by ouabain.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1424
    Keywords: Ouabain-resistance ; Epithelia ; Ma104 cells ; MDCK cells ; Tight junctions ; Na+, K+-ATPase ; K+ content ; Metabolic cooperation ; Cell attachment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Ma104 cells (renal, epithelial) have a peculiar way of resisting ouabain: their Na+,K+-pumps bind the drug with high affinity, cellular K+ is lost and cell division arrested, but cells do not detach as most cell types do. Then, if up to 4 days later the drug is removed, Ma104 cells recover K+ and resume proliferation (Contreras et al., 1994). In the present work, we investigate whether Ma104 cells are able to protect ouabain-sensitive MDCK cells in co-culture. The main finding is that they do, but in this case protection is not elicited by the usual mechanism of maintaining the K+ content of neighboring cells through cell-cell communications. Ma104 cells treated with ouabain simply remain attached to the substrate and to their MDCK neighbors, and both cells lose K+. This attachment includes tight junctions, because the transepithelial electrical resistance of the monolayers is not abolished by ouabain. Although the β-subunit of the Na+,K+-ATPase is known to possess molecular characteristics of cell-cell attachment molecules, attachment between Ma104-MDCK cells does not seem to be mediated by this enzyme, as immunofluorescence analysis reveals that Na+,K+-ATPase is only inserted in the plasma membrane facing a neighboring cell of the same type.
    Type of Medium: Electronic Resource
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